Psoralen were evaluated from in vivo and in vitro plant parts of Psoralea corylifolia by High Performance Liquid Chromatography (HPLC). The high amount of psoralen (3038 μg/g fresh weight) in in vivo condition were obtained in brown seeds. Psoralen content in plant parts growing in vitro were lower than those growing in vivo. Quantitative analysis on psoralen in callus derived from different plant parts showed that maximum psoralen (2601.8 μg/g fw) was recorded in callus derived from cotyledons. Estimation of psoralen in shoots derived from different callus were indicated high amount of psoralen (572.3 μg/g fw) in plant drivated from node callus. Identification of psoralen in the callus of Psoralea corylifolia is reported for first time. In the other section attempts were done to enhance of psoralen by some organic elicitors including; yeast extract, proline, myzo-inositol and sucrose.in different medium. For the first time a comparative study of mature nodal, juvenile nodal explants and cotyledonary callus cultures were evaluated. These explants elicitated with different organic elicitor. Results revealed that maximum quantity of psoralen were present in cotyledonary callus cultures, mature and juvenile nodal cultures, respectively. Also, results indicated that there were significant variations in the psoralen content in mature nodal, juvenile nodal and cotyledonary callus cultures, when their medium supplemented with different elicitors in the wide range of concentrations. Interestingly, high amount of psoralen content (2881.9 μg/g fw.) even among all of the organic elicitors tried was found at 300 mg/l yeast extract in cotyledonary callus cultures.